Insights into the Biocompatibility and Biological Potential of a Chitosan Nanoencapsulated Textile Dye.

Traditionally synthetic textile dyes are hazardous and toxic compounds devoid of any biological activity. As nanoencapsulation of yellow everzol textile dye with chitosan has been shown to produce biocompatible nanoparticles which were still capable of dyeing textiles, this work aims to further characterize the biocompatibility of yellow everzol nanoparticles (NPs) and to ascertain if the produced nanoencapsulated dyes possess any biological activity against various skin pathogens in vitro assays and in a cell infection model. The results showed that the NPs had no deleterious effects on the HaCat cells' metabolism and cell wall, contrary to the high toxicity of the dye. The biological activity evaluation showed that NPs had a significant antimicrobial activity, with low MICs (0.5-2 mg/mL) and MBCs (1-3 mg/mL) being registered. Additionally, NPs inhibited biofilm formation of all tested microorganisms (inhibitions between 30 and 87%) and biofilm quorum sensing. Lastly, the dye NPs were effective in managing MRSA infection of HaCat cells as they significantly reduced intracellular and extracellular bacterial counts.

Conventional and natural compounds for the treatment of dermatophytosis.

Dermatophytes are a group of pathogenic fungi that exclusively infect the stratum corneum of the skin, nails, and hair, causing dermatophytosis. Superficial skin infections caused by dermatophytes have increased in the last decades. There are conventional antifungals that treat these infections, such as terbinafine, fluconazole, and others. However, the limitations of these treatments (resistance, side effects and toxicity) along with the increasing over-prescription, the misuse of these antifungals and the high treatment costs led to the search for new, alternative, natural-based antifungal drugs. These have multiple mechanisms of action, which works to their advantage, making it difficult for a fungus to create resistance mechanisms against all of them at the same time. The main objective of this work is to provide a state-of-the-art review on dermatophytes, dermatophytosis, and the existing treatments, both conventional and natural, such as chitosan and essential oils.

Chitosan's biological activity upon skin-related microorganisms and its potential textile applications.

Over the years the body of work relating chitosan and its possible skin-related applications has grown, showing that chitosan is active both as a free compound and as a functional finishing of textiles. As a free molecule chitosan proved itself to be an attractive option as it is biocompatible and has a demonstrated biological activity (e.g. growth inhibition or adhesion inhibition) upon multiple skin pathogens, even upon multidrug resistant species. Furthermore, chitosan has wound healing accelerating properties, which make it a prime candidate for possible control of skin related infections. Almost inconspicuously, textiles have been one the main lines of defense of the skin against foreign threats, acting as a physical barrier to microbial colonization and infection. With the advent of textile functionalization specially designed textiles with enhanced protective characteristics, such as antimicrobial capacity, have come to the forefront. Chitosan functionalized textiles have been shown to be effective inhibitors of microbial growth with even invasive multidrug resistant species, as MRSA, being inhibited. Therefore, chitosan and chitosan functionalized textiles present themselves as both an interesting alternative to traditional antibiotics and as a possible means to enhance current treatment strategies.

Plant-Based Films and Hydrogels for Wound Healing.

Skin is constantly exposed to injury and infectious agents that can compromise its structural integrity and cause wounds. When this occurs, microorganisms from the skin microbiota and external bacteria and fungi can penetrate the wound and cause an infection, which complicates the healing process. Nowadays, there are several types of wound dressings available to treat wounds, some of which are incorporated with antimicrobial agents. However, the number of microorganisms resistant to these substances is rising. Therefore, the search for new, natural alternatives such as essential oils (EOs) and plant extracts (PEs) is on the rise. However, these substances present some limitations (poor bioavailability and poor target capacity), which limits their efficiency. Their incorporation in formulations in the form of films and hydrogels (HGs) can help to overcome these issues and may be a potential alternative to the current treatments. HGs and films incorporated with PEs and EOs have antimicrobial activity, promote the viability of skin cells and fibroblast migration, and are non-toxic and biocompatible. This review discusses the use of films and HGs for the topical delivery of EOs and PEs for wound treatment and their formulations as effective wound dressings, while debating some mechanisms and biological properties to elucidate their presumptive clinical relevance and possible applications.

An Overview on Mushroom Polysaccharides: Health-promoting Properties, Prebiotic and Gut Microbiota Modulation Effects and Structure-function Correlation.

Mushroom polysaccharides are recognized as "biological response modifiers". Besides several bioactivities, a growing interest in their prebiotic potential has been raised due to the gut microbiota modulation potential. This review comprehensively summarizes mushroom polysaccharides' biological properties, structure-function relationship, and underlying mechanisms. It provides a recent overview of the key findings in the field (2018-2024). Key findings and limitations on structure-function correlation are discussed. Although most studies focus on ß-glucans or extracts, α-glucans and chitin have gained interest. Prebiotic capacity has been associated with α-glucans and chitin, while antimicrobial and wound healing potential is attributed to chitin. However, further research is of utmost importance. Human fecal fermentation is the most reported approach to assess prebiotic potential, indicating impacts on intestinal biological, mechanical, chemical and immunological barriers. Gut microbiota dysbiosis has been directly connected with intestinal, cardiovascular, metabolic, and neurological diseases. Concerning gut microbiota modulation, animal experiments have suggested proinflammatory cytokines reduction and redox balance re-establishment. Most literature focused on the anticancer and immunomodulatory potential. However, anti-inflammatory, antimicrobial, antiviral, antidiabetic, hypocholesterolemic, antilipidemic, antioxidant, and neuroprotective properties are discussed. A significant overview of the gaps and research directions in synergistic effects, underlying mechanisms, structure-function correlation, clinical trials and scientific data is also given.

Novel Strategies for Preventing Dysbiosis in the Oral Cavity.

Oral diseases affect over three billion people worldwide, making it one of the most common infections. Recent studies show that one approach to reducing the risk of chronic infections, such as caries, gingivitis, periodontitis, and halitosis, is to control the ecology of the oral microbiome instead of completely removing both the harmful and beneficial microorganisms. This is based on the knowledge that oral diseases are not caused by a single pathogen but rather by a shift in the homeostasis of the entire microbiota, a process known as dysbiosis. Consequently, it is of the utmost importance to implement strategies that are able to prevent and control oral dysbiosis to avoid serious complications, including heart, lung, and other systemic diseases. Conventional treatments include the use of antibiotics, which further disrupt the equilibrium in the oral microbiota, together with the mechanical removal of the decayed cavity area following its formation. Therefore, it is imperative to implement alternative strategies with the potential to overcome the disadvantages of the current therapy, namely, the use of broad-spectrum antibiotics. In this sense, probiotics and postbiotics have received particular attention since they can modulate the oral microbiota and decrease the dysbiotic rate in the oral cavity. However, their mechanisms of action need to be addressed to clarify and drive their possible applications as preventive strategies. In this sense, this review provides an overview of the potential of probiotics and postbiotics, focusing on their antimicrobial and antibiofilm activities as well as their ability to modulate the inflammatory response. Finally, it also showcases the main advantages and disadvantages of orodispersible films-a promising delivery mechanism for both probiotics and postbiotics to target oral dysbiosis.

Impact of LAB from Serpa PDO Cheese in Cheese Models: Towards the Development of an Autochthonous Starter Culture.

Serpa is a protected designation of origin (PDO) cheese produced with raw ewes' milk and coagulated with Cynara cardunculus. Legislation does not allow for milk pasteurization and starter culture inoculation. Although natural Serpa's rich microbiota allows for the development of a unique organoleptic profile, it also suggests high heterogeneity. This raises issues in the final sensory and safety properties, leading to several losses in the sector. A possible solution to overcoming these issues is the development of an autochthonous starter culture. In the present work, some Serpa cheese Lactic acid bacteria (LAB)-isolated microorganisms, previously selected based on their safety, technological and protective performance, were tested in laboratory-scale cheeses. Their acidification, proteolysis (protein and peptide profile, nitrogen fractions, free amino acids (FAA)), and volatiles generation (volatile fatty acids (VFA) and esters) potential was investigated. Significant differences were found in all parameters analyzed, showing a considerable strain effect. Successive statistical analyses were performed to compare cheese models and Serpa PDO cheese. The strains L. plantarum PL1 and PL2 and the PL1 and L. paracasei PC mix were selected as the most promising, resulting in a closer lipolytic and proteolytic profile of Serpa PDO cheese. In future work, these inocula will be produced at a pilot scale and tested at the cheese level to validate their application.

Chitosan Nanoparticles as Bioactive Vehicles for Textile Dyeing: A Proof of Concept.

In recent years bioactive textiles have risen to the forefront of consumers perception due to their potential protection against virus, fungi and bacteria. However, traditional textile staining is an eco-damaging process that and current methods of textile functionalization are expensive, complicated and with great environmental impact. With that in mind, this work sought to show a possible solution for this problematic through the usage of a novel one step textile dyeing and functionalization method based upon nanoencapsulated textile dyes (NTDs). To do so navy blue everzol NTDs were produced with chitosan, cotton dyed, characterized through FTIR and SEM and biological potential evaluated through biocompatibility screening and antimicrobial activity against skin pathogens. The data obtained showed that NTDs effectively dyed the target textile through a coating of the cotton fibre and that NTDs formed hydrogen bonds with the cellulose fibre via electrostatic interactions of the chitosan amino groups with cotton sulphate groups. From a biocompatibility perspective NTDs dyed cotton had no deleterious effects upon a skin cell line, as it promoted cellular metabolism of HaCat cells, while traditionally died cotton reduced it by 10%. Last but not least, NTDs dyed cotton showed significant antimicrobial activity as it reduced viable counts of MRSA, MSSA and A. baumannii between 1 and 2 log of CFU while traditional dyed cotton had no antimicrobial activity. Considering these results the novel method proposed shows is a viable and ecological alternative for the development of antimicrobial textiles with potential biomedical applications.

Organoleptic Chemical Markers of Serpa PDO Cheese Specificity.

Serpa is a protected designation of origin cheese produced with a vegetable coagulant (Cynara cardunculus L.) and raw ovine milk. Despite the unique sensory profile of raw milk cheeses, numerous parameters influence their sensory properties and safety. To protect the Serpa cheese quality and contribute to unifying their distinctive features, some rheologic and physicochemical parameters of cheeses from four PDO producers, in distinct seasons and with different sensory scores, were monitored. The results suggested a high chemical diversity and variation according to the dairy, month and season, which corroborates the significant heterogeneity. However, a higher incidence of some compounds was found: a group of free amino acids (Glu, Ala, Leu, Val and Phe), lactic and acetic acids, some volatile fatty acids (e.g., iC4, iC5, C6 and C12) and esters (e.g., ethyl butanoate, decanoate and dodecanoate). Through the successive statistical analysis, 13 variables were selected as chemical markers of Serpa cheese specificity: C3, C4, iC5, C12, Tyr, Trp, Ile, 2-undecanone, ethyl isovalerate, moisture content on a fat-free basis, the nitrogen-fractions (maturation index and non-protein and total nitrogen ratio) and G' 1 Hz. These sensory markers' identification will be essential to guide the selection and development of an autochthonous starter culture to improve cheese quality and safety issues and maintain some of the Serpa authenticity.

Textile dyes loaded chitosan nanoparticles: Characterization, biocompatibility and staining capacity.

Textile dyeing is a hazardous and toxic process. While traditionally it has been managed through effluent treatment, new approaches focused upon improving the dyeing process are gaining relevance. In this work, we sought to obtain, for the first time, an eco-friendly chitosan-nanoparticle based textile dyeing method. To that end, yellow everzol and navy blue itosperse loaded chitosan nanoparticles were produced and their capacity to dye textiles and cytotoxicity towards human skin cells were evaluated. The results obtained showed that it was possible to obtain nanoencapsulated dyes through ionic gelation with an average entrapment efficacy above 90 %. Nanoparticles presented a positive surface charge and sizes between 190 and 800 nm with yellow everzol NPs occurring via ionic interactions while navy blue itosperse NPs were formed through hydrogen bonds. Furthermore, the produced dye NPs presented no cytotoxicity towards HaCat cells and presented staining percentages reaching 17.60 % for a viscose/wool blend.

In vitro screening for anti-microbial activity of chitosans and chitooligosaccharides, aiming at potential uses in functional textiles.

Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) skin disorders. Hence, this research effort was aimed at elucidating the relationship between molecular weight (MW) of chitosan and its antimicrobial activity upon six dermal reference microorganisms, as well as the influence of the interactions with cotton fabrics on said activity. Using 3 chitosans with different MW, as well as two chitooligosaccharide (COS) mixtures, a relevant antimicrobial effect was observed by 24 h for the six microorganisms tested; it was apparent that the antimicrobial effect is strongly dependent on the type of target microorganism and on the MW of chitosan being higher for lower MW in the case of E. coli, K. pneumoniae and P. aeruginosa, and the reverse in the case of both Gram-positive bacteria. Furthermore, a strong anti-fungal effect was detectable upon C. albicans, resembling the action over Gram-positive bacteria. Interactions with cotton fabric resulted in a loss of COS activity when compared with cultured media, relative to the effect over Gram-negative bacteria. However, no significant differences for the efficacy of all the 5 compounds were observed by 4 h. The three chitosans possessed a higher antimicrobial activity when impregnated onto the fabric, and presented a similar effect on both Gram-positive bacteria and yeast, in either matrix. Pseudomonas aeruginosa showed to be the most resistant microorganism to all five compounds.

Antimicrobial effects of chitosans and chitooligosaccharides, upon Staphylococcus aureus and Escherichia coli, in food model systems.

The objective of this study was to elucidate the controversial relationship between the molecular weight (MW) of chitosans and their antibacterial activity (upon different inoculum levels, at several concentrations). The influence of food components on the activity was also ascertained, as well as acceptance by a sensory panel. All the compounds tested exhibited antibacterial activity against Staphylococcus aureus and Escherichia coli. This activity was shown to be closely dependent on the inoculum level, MW and concentration used. Within 4h at 10(3) cells/mL, all five compounds, at every concentration (0.5%, 0.25% and 0.1%, w/v), proved to be bactericidal; for higher inocula, 0.1% (w/v) was only bacteriostatic; at 10(7) or 10(5) cells/mL, and independently of the inoculum level, 0.25% (w/v) of any chitooligosaccharide (COS) mixture was sufficient to reduce the E. coli initial population by at least 3 log cycles; COS never exhibited bactericidal action over S. aureus, unlike high and medium MW chitosans-which, at 0.5% (w/v), presented a bactericidal effect even against 10(7) cells/mL. When incorporated in liquid food matrices, medium and high MW chitosans maintained their activity, for both matrices and bacteria, although a slower activity was noticeable in milk; however, COS lost their activity upon both bacteria in milk after 4-8h. Furthermore, addition of chitosans to apple juice led to several unpleasant off-flavors, such as astringency and after taste--which increased in magnitude with MW.

Exploring chitosan nanoparticles as effective inhibitors of antibiotic resistant skin microorganisms - From in vitro to ex vitro testing.

Nowadays, nosocomial skin infections are increasingly harder to manage and control. In the search for new, natural compounds capable of being alternatives to traditional antibiotics, chitosan and its nanoparticles, have garnered attention. This work sought to understand the potential of chitosan NPs in the management of infections caused by MDR skin pathogens in planktonic and sessile assays. Additionally, NPs' capacity to inhibit biofilm quorum sensing and prevent HaCat infections was also evaluated. The results obtained showed that chitosan NPs had an average size and charge of 226.6 ± 5.24 nm and +27.1 ± 3.09 mV. Inhibitory and bactericidal concentrations varied between 1 and 2 mg/mL and 2-7 mg/mL, respectively. Chitosan NPs effectively inhibited biofilm growth for all microorganisms and possessed strong anti-quorum sensing activity. Lastly, chitosan NPs proved to be effective interfere with A. baumannii's infection of HaCat cells, as they significantly reduced intracellular and extracellular bacterial counts.

Investigation of chitosan's antibacterial activity against vancomycin resistant microorganisms and their biofilms.

Vancomycin-resistant microorganisms are a hurdle that traditional antibiotics struggle to overcome. These difficulties have led to search for new solutions based on natural products. Chitosan has been recognized as an effective antibacterial agent against a vast array of microorganisms including antibiotic resistant ones. As such, this work aimed to evaluate chitosan as an alternative to traditional antibiotics in the management/control of two vancomycin-resistant microorganisms, VRSA and VREF, in planktonic and sessile settings. The results obtained showed that chitosan was highly effective in inhibiting VRSA and VREF planktonic growth and reduced VREF viable counts by 6 log CFU in 30min. Additionally, chitosan was active upon several phases of VRSA and VREF sessile growth inhibiting adhesion, biofilm formation and dual-species biofilms at concentrations as low as 0.0125mg/mL. In lieu of these results chitosan shows great potential as a possible alternative for the control of vancomycin-resistant microorganisms in recalcitrant wound infections.

Assessment of the prebiotic effect of quinoa and amaranth in the human intestinal ecosystem.

Quinoa and amaranth belong to the group of the so called "superfoods" and have a nutritional composition that confers multiple benefits. In this work, we explored the possibility of these foods exhibiting a prebiotic effect. These pseudocereals were subjected to an in vitro digestion and used as carbon sources in batch cultures with faecal human inocula. The effects on the microbiota composition and their metabolic products were determined by assessment of variations in pH, short-chain fatty acid (SCFA) production and changes in the dynamic bacterial populations by fluorescence in situ hybridization (FISH). After 48 h of incubation, the total SCFAs were 106.5 mM for quinoa and 108.83 mM for amaranth, in line with the decrease in pH. Considerable differences (p < 0.05) were found in certain microbial groups, including Bifidobacterium spp., Lactobacillus-Enterococcus, Atopobium, Bacteroides-Prevotella, Clostridium coccoides-Eubacterium rectale, Faecalibacterium prausnitzii and Roseburia intestinalis. Our research suggests that these pseudocereals can have the prebiotic potential and that their intake may improve dysbiosis or maintain the gastrointestinal health through a balanced intestinal microbiota, although additional studies are necessary.

Influence of abiotic factors on the antimicrobial activity of chitosan.

In an effort to bypass the adverse secondary effects attributed to the traditional therapeutic approaches used to treat skin disorders (such as atopic dermatitis), alternative antimicrobials have recently been suggested. One such antimicrobial is chitosan, owing to the already proved biological properties associated with its use. However, the influence of abiotic factors on such activities warrants evaluation. This research effort assessed the antimicrobial activity of chitosan upon skin microorganisms (Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli) in vitro when subject to a combination of different abiotic factors such as pH, ionic strength, organic acids and free fatty acids. Free fatty acids, ionic strength and pH significantly affected chitosan's capability of reducing the viable numbers of S. aureus. This antimicrobial action was potentiated in the presence of palmitic acid and a lower ionic strength (0.2% NaCl), while a higher ionic strength (0.4% NaCl) favored chitosan's action upon the reduction of viable numbers of S. epidermidis and E. coli. Although further studies are needed, these preliminary results advocate that chitosan can in the future be potentially considered as an antimicrobial of choice when handling symptoms associated with atopic dermatitis.

Study of antimicrobial activity and atomic force microscopy imaging of the action mechanism of cashew tree gum.

The aim of this work was to evaluate the antimicrobial potential of two grades of cashew tree gum (crude and purified) against eight microorganisms and to analyze the mechanism of cashew tree gum antimicrobial action via atomic force microscopy (AFM) imaging. The results indicated strong antimicrobial properties of pure cashew tree gum against all tested microorganisms, except for Candida albicans and Lactobacillus acidophilus. On the other hand crude cashew gum showed antimicrobial activity only against Gram-positive bacteria (MRSA, MSSA, Listeria innocua and Enterococcus faecium). Atomic force microscopy imaging showed that pure cashew tree gum lead to bacterial cell collapse. In conclusion cashew tree gum presented relevant antimicrobial activity against most of the studied bacteria, and the purification of the cashew gum affected its antimicrobial spectrum.

Biodiversity and characterization of Staphylococcus species isolated from a small manufacturing dairy plant in Portugal.

The level and the diversity of the staphylococcal community occurring in the environment and dairy products of a small manufacturing dairy plant were investigated. Species identification was performed using different molecular methods, viz. Multiplex-PCR, amplified ribosomal DNA restriction analysis (ARDRA), and sodA gene sequencing. The main species encountered corresponded to Staphylococcus equorum (41 isolates, 39.0%), S. saprophyticus (28 isolates, 26.7%) and S. epidermidis (15 isolates, 14.3%). Additionally, low incidence of enterotoxin genes was obtained, with only 9 strains (8.6%) being positive for one or more toxin genes. With regard to antimicrobial resistance, 57.1% of the isolates showed at least resistance against one antibiotic, and 28.6% were multi-resistant, which might accomplish resistance for up to 6 antibiotics simultaneously. These results provided evidence that the presence of Staphylococcus species in dairy environment are mostly represented by S. equorum and S. saprophyticus, and illustrate that carrying antimicrobial resistance genes has become reasonably widespread in cheese and dairy environment.